PERSOONIA
Volume
two Neottiella species
Bellis
Estonian
Genome
life
size,
fruit-bodies
two
(Fr.)
ploidy
conidia
of
to have
level
of
genomes
of N.
content of 60
Mb,
54
The two species
their
in
ascospore
are
(25 Mb).
N.
of
reveals
rutilans,
the
vivida
ascus
2C value but
of
Quantification
understanding
changes
al., 1983;Horgenetal.,
Changes
species,
cycle
half
at
content
In the
development
varies
identical in
of
and
The
term
is known
can
were
In N.
with
found
ploidy
The
vivida,
a
DNA
distinguished only
and waited in
of N.
vivida,
rutilans, endoreduplication
at a different rate
levels
be
N. rutilans
ascospores
of
sporal
may
in
spores.
nuclei.
In
result from
ploidy-dependent
content
Thus N.
N.
ruti-
vivida
different
the
is not
and
patterns
gene.
contributed
significantly
to
better
a
fungi (Olive, 1953; Bryant & Howard, 1969;
1982; Collins
occur at
ploidy
et
as
during
of the
the mitotic (G1
terms
and mitosis.
al. 1983; Whisler
et
an
and cell division, in the
organism. Ascomycetes
and reduce their
ascus
phases) cycle
cell division. Cells
cell
and
is
a
arise
are
often
through
their
differing only by
physiological
content
haploid
of the
(Swift, 1950). Dividing
as
of cell division.
types may
quite
characteristics
ploidy
different in
(Galitski
et
1999).
The DNA
known
Interphase
G2
of DNA sequence information, but
quantity.
events
versus
organism, specialized polyploid
cycles lacking
its lC-value
sequence of
replication
upon fertilization in the
of the
genome denotes the DNA
and
DNA
differentiation of cells in
developmental, morphological,
quality
In N.
1983; Anderson,
during
al., 1999; Hieter & Griffiths,
of
70.
with
spore-
1985; Bresinsky etal., 1987; Wittmann-Meixner, 1989;Bayman
double their
i.e. cell
endocycles,
terms
al.,
to be
a
meiosis, producing ascospores. In the vegetative mycelium, the nuclear
DNA
are
similar and
in nuclear DNA has
of nuclear DNA
sexual
a
et
and N. rutilans
is reticulated in
ornamentation
the
and
1990; Weber, 1992).
evolution of
ploidy by
spore
(Nyl.)
by comparison
mitochondria
giant
the uninucleate
ploidy
of the life cycle of several
Collins, 1979; Franklin
& Collins,
in
expression regulated by
gene
vivida
measured from
found.
very
proceed
may
heterogeneity
in
was
Mb, respectively.
calculated
was
were
morphologically
530
where
apex
ornamentation, which
differences
obtained
Neottiella vivida
of their nuclear DNA content is 2C.
arrested at the
lans
was
750 Mb and
N. vivida. Due to endoreduplication in
value
DNA content
size
genome
and 30 Mb
Mb,
Botany,
determined for Neottiella
were
that of N.
50,
occurs
and
Zoology
Trichophea hemisphaerioides (23.3 Mb)
approximately
rutilans is
meiotic division
Ascomycota)
e-mail: [email protected]
Dennis. The relative
from Pleurotus ostreatus
print
by
and
cycle,
using cytofluorometry;
standards:
Institute of
St., 51014Tartu, Estonia;
Dennis and N. rutilans
and ploidy
(Pezizales,
Kullman
Agricultural University,
181 Riia
with
life cycle
content,
Nuclear DNA
in
18, Part 1, 103-114(2002)
the cell
period
cycle.
chromosome complement in
unreplicated haploid
cells pass
The cell
of chromosome
cycle
through
is
a
terms
nuclear genome
regular, repeated
divided into
duplication. Interphase
interphase
can
be divid-
104
PERSOONIA
ed into three phases, which
are
referred
is
state
called the GO
controlled
the
cycle
At the G2
mitosis
triggers
or
et
al.,
of
Saccharomyces
cerevisiae
or
metaphase,
nuclei of spores of most
the
mitosis is
cycle
can
distribution
of the
nuclei in the
a
In the
unit).
al.,
1995)
are
cannot
cycle
et
1992),
and
large-budded
al.,
species
2000)
anaphase
the
1995). However,
well
as
spores
fraction
as
Glomus versi-
in the genera Pleuro-
arrested in the G0/G1
are
reveal whether each individual nucleus
not, kinetic information can be inferred
or
cycle). Progression
through
The
content.
haplophase (nuclei
GO/G1
phase, corresponds
with
the
position
tested
was
in
same
as
S-phase
comparable
cycle.
In
et
to
al.
haplonts
(1C-value
the DNA
content
Wittman-Meixner &
(1987),
due
to
Bresinsky
It was shown that basic nuclear
mycelia (irrespective
or
of the medium and
conidia. In young
Most nuclei
fruit-body.
nuclear DNA
indicating
-
unit) (see Kullman, 2000).
in fruit-bodies, sclerotia
those of the
content.
set' of chromosomes),
DNA content),
(basic
Bresinsky (1992).
in young and old
as
'single
and
of the nuclei in
variation of nuclear relative DNA
fungi,
by Bresinsky
well
a
genome size
to
nuclear genome,
dikaryophase,
two
nuclei combine
'double set' of chromosomes).
the nucleus is
ascus,
the
haploid (n)
mitotic cell
(x
division)
-
and
divided
In
fungi
are
mycelia,
in the G0/G1
content
in this cell
to
form
zygote (nuclei in diplophase
the
a
zygote
immediately by
condition in the life
The cell nuclei may
to occur
initiates
cycle phase
corre-
their genome size.
to
restoring
et
is obtained whose first maximum
cycle
is the
of the cell
In the
with
or
arrested in the
are
cycle (Hanna
in nuclear DNA
Weber (1992), and Weber &
content
sponds
per se
of the cell
(position
the age of the culture),
phase
cycle
often have uninucleate
Reese) Hansen
Quel. (Kullman,
the cell
through
polyploidy
cycle
DNA values
ex
of the cell
unreplicated haploid
In studies of
DNA
the
therefore be estimated on the basis of measurement of DNA
curve
nuclei in the cell
(1989),
phase (synthesis phase).
arrests
(Helotiales)
in Helotiales (Weber,
measurements
content
the mitotic
mainly
the end of G1
that the control system either
checkpoint
The undivided nuclei of the
(Bianciotto
expressed by changes
measuring
content
is
1C-value).
progresses
from DNA
a
species
S.M. Berch
Hence, although
actually
one at
-
dormant,
or
cycle, progression
checkpoints
either
system again
(Meyen
phase
P. Kumm. and Phellinus
phase (at
When
1992).
i.e. in the G2
forme (R Karst.)
In the cell
1999).
(Weber,
the cell
process
cytokinesis together
specialized resting,
that will initiate the S
in Sclerotiniaceaeand Leotiaceae
Species
(Fr.)
the G1
at
2002
1,
The
called
points,
the control
checkpoint,
(Raven
a
Part
and G2. Mitosis and
cycle.
phase (G-zero phase).
with the 2C-value
tus
of the cell
the end of G2. It is
at
18,
designed G1, S,
are
phase
crucial transition
at two
and another
arrests
the M
to as
Vol.
cycle (n
undergo endoreduplication
endopolyploidy ('many
is the
meiosis
-
the
only diploid (2n)
cell.
(zygotic meiosis),
thus
haploid
chromosome set,
(DNA replication in absence of
set' of chromosomes)
the basic chromosome set, thebasic DNA
content
in
can
be assumed
germ-line polyploids)
(Nagel, 1978).
Since in
paraphyses
assumed
Serpula,
,
some cases
the
content
but also in all other cell
to
and
occur
in
ascomycetes
of nuclear DNA in the
types) appeared
(Weber,
1992)
to
be
and in
fruit-body (often
in
tips
larger, endopolyploidy
basidiomycetes
(Paxillus,
Leucogyrophana) (Meixner & Bresinsky, 1988; Wittman-Meixner&
sinsky, 1989).
of
was
Bre-
Kullman:
In this
Nucleair
DNA
the nuclear behaviour of
study
in
two Neottiella
parasites,
two moss
vivida and N. rutilans, is examined. The main aim
in
changes
and
ploidy during fungal growth
absolute genome sizes and
MATERIALS
Fruit-bodies ofNeottiella vivida
1998, A. Jakobson (TAA
morphogenesis,
levels in these
ploidy
kept
at
form: l
measuring
and N.
x
w
mean
rutilans (Fr.)
fixed in
were
26
Aug.
Dennis, Finland, Kilpisjarvi,
Carnoy (Romeis, 1948)
and
the
to
w
mean
a
content were
are
denote the
mean
percentage divided by the
mean
mea-
HI
an
values of the
length
equivalent
to
in nuclei and mitochondria was measured
Botany, Regensburg University, using
ped
with
meter.
to
III RS
described in
Bresinsky
to
the
et
cover-slip
al.
(1987),
genome
in nuclear DNA
relative DNA
content
content
is
population
by multiplying
expressed
nucleotide
Mb,
see
The
two
in
is converted
in
the
the result
the
Bennet & Leitch,
genome
standards:
to
of nuclei by
DNA)
size of N.
by
and
can
the
mean
microscope photo-
a.u.) is proportional
haplophase,
nuclei in the cell
analyzed
for
cycle
a
distribution
G0/G1
calculating
mean
amount.
an
phase,
content
size of the standard.
pairs (bp),
megabase pairs
the difference
internal standard,
The genome size of an
relative DNA
of the standard G0/G1
in base
or
be
the absolute
genome
vivida and N.
a
kilobases
of nucleotides
population
(kb,
a
un-
of the unknown
Usually
of nuclei
DNA
content
stretch of
(Mb) (NB
rutilans
was
estimated
pure culture of the ascomycete
Graddon
(TFC
from the oyster mushroom Pleurotus
The genome size of T.
=
lpg
1000
=
965
1995).
conidia from
sphaerioides (Mounton)
units
specimens. By including
dividing
picograms (pg)
pairs
con-
the Institute of
photomicroscope, equip-
03 Zeiss
nuclei in the
indicating
histograms
between the
known specimen is obtained by
G0/G1
an
at
size.
The resulting fluorescence
and
measuring
is obtained whose first maximum,
corresponds
Zeiss UNIVERSAL
intensity (in arbitrary
in the nucleus. When
content
a
by cytofluorometry
illuminator, and
epifluorescence
The measured fluorescence
DNA
curve
an
the
value.
Wittmann-Meixner (1989), Weber (1992), and Biittner (1999). The relative DNA
tent
100
in the follow-
between the slide and the
squeezed
DAPI-staining procedure
mean
with
presented
The variation coefficient is
specimen.
was
also used for
microscope equipped
and width (w) of spores
nuclei the material
staining
subjected
a
'AMPLIVAL'
where l
pm,
mean
standard deviation as
For
occur
determine
to
species.
relative nuclear DNA
an
length (1)
and width of 20 spores from
and
as
(Nyl.) Dennis, Norway, Tromso, Brennfjell,
135733)
spore dimensions with
immersion objective. The
ing
well
4°C until needed.
The slides used for
suring
as
AND METHODS
Sana, 25 Aug. 1998, A. Jakobson (TAA 135730),
and
the ascomycetes Neottiella
establish whether there
to
was
105
species
97-71 from TAA
ostreatus
hemisphaerioides
was
by comparison
Trichophaea
147708)
and
a
with
hemi-
spore-print
(Jacq.: Fr.) R Kumm. (TAA 142824).
23.3 Mb, and that of P.
ostreatus
25 Mb
(Kullman, 2000).
RESULTS AND
From the
typical
standpoint
of nuclear
of ascomycetes (Rossen &
cytology,
DISCUSSION
N. vivida and N. rutilans have
Westergaard, 1966; Weber, 1992;
a
life
cycle
Weber & Bresins-
106
Fig.
a.
c.
PERSOONIA
1.
Diagram representation
Crozier formation from
tip
first
and basal
one.
nuclei
Nuclei
in
between
Fig.
2.
Fig.
3.
(ascus
h.
two
e.
oil
mother cell
to
j.
Uninucleate
conjugate nuclear
a
g.
young
i.
(zygote
after the
ascospore
ascospore.
in N. vivida.
ascus
mitotic division
another crozier
ascus
four nuclei
(Mi), 1C;
mature
or
the
by
nucleus
second
the side
=
4C);
meiotic
(Mi);
of the
f.
two
division
formation; endoreduplication
Nucleus
with
condensed chromatin
globules.
Synchronous
up
(Mel), 2C;
b.
mother cell
in
karyogamy (K)
nuclei after mitotic division
ascospores;
Anaphase nuclei
ky, 1992).
a
eight
young
young ascus,
nuclei,
2C; d,
to
and form ascus
after the first meiotic division
(Me2), 1C;
(ERe)
up
fuse
1, 2002
during developmentof
hypha;
ascogenous
Pan
18,
of nuclear behaviour
dikaryotic
crozier cells
Vol.
with
DNA
is not
d. four
4C;
unreplicated DNA
synthesis
in
demonstrated), a,
products
content
all crozier
b. DNA
of meiosis each
cells
(Mi)
in
haplontic hyphae
before meiosis
replication,
approaching
nuclei
up
in
to
the
2C;
mitotic division
of N. vivida.
of N.
ascus
c.
vivida
karyogamy
in
(DNA synthesis
a
in
2C).
The
cycle
dikaryophase,
includes
a
monokaryotic
thallus (with nuclei in the
haplophase),
confined upon fructification, and asci which represent zygotes.
the nuclei are in the
sion is meiotic and
diplophase
meiospores
of the life cycle. When the
are
formed (Figs.
ascus
Here
nucleus divides, divi-
1, 3,4a-h).
Somatic nuclear behaviour
The presence of up
indicates mitotic cell
of a
at
fruit-body
to
double DNA
cycle phases
nuclei divide within the
maturity (except
content
in the nuclei of
G0/G1 and G2/M
hyphae
hyphae
and
but remain stable within the
for the nuclei of their apical
paraphyses
(Figs. 2,6d, e). During
cells).
the
growth
paraphyses
Kullman:
vivida,
In N.
of the nuclei
nuclei of the
the G0/G1
some
more
the
tip
content
while in the second
in the G2/M
The
phase.
variable compared with the
When
fluorescence
measuring
are
used, and 12% when spores
of nuclear DNA-DAPI
preferred
of the
paraphyses
paraphyses
and
suitable for
measure-
For this
noise).
too,
reason
and
content
auto-
used, 12% when hyphae
are
determinedas 750 Mb
was
paraphyses
are more
phase
larger
due to the mitot-
used from total measured fluorescence
+
is
hyphae
in
and
subhymenium hyphae.
in determinationof relative DNA
1C-value, i.e. genome size,
of the
only
nuclei, noise (additional light including the
on
are
complex
content
750 Mb ± 9% and in the
the measured nuclei were
content
content
for 6% when
accounts
(DNA
paraphyses
was
nuclei were in the G0/G1
DNA
DNA
of the C-value than the nuclei of the
fluorescence of cells)
case
case, most
mean
mean
case)
107
species
in the nuclei of the
ic division of the first. Hence the nuclei of the
ment
two Neottiella
in
measured in this
not
was
DNA
770 Mb ± 36%. In the first
hyphae
phase,
were
DNA
mean
the
at
Nucleair
(fluorescence
should be
paraphyses
size. In this
genome
case
for,N.
530 Mb
vivida and
as
the
for N. rutilans (see Materials and Methods).
Development of the
The asci of.N.
of
from croziers
develop
(Read & Beckett,
karyogamy
the hooked cell
ating
ascus
vivida
undergo conjugate
three cells
(Fig.
la-c).
according
1996; Chiu & Moore,
to
the Neottiella-type pattern
1999) (Fig. 1).
mitoses (Mi) after which
two
The three cells of the crozier
septa
Two nuclei in
formed
are
cre-
termed the terminal
are
cell, the penultimate cell, and the stalk cell, representing respectively the first, the second,
and the third cell of the crozier. The
other cells
are
are
uninucleate. At first,
penultimate
two
prefusion
cell is
located in the terminal cell and in the stalk cell,
terminal cell then
non-sister nuclei, may become the
taining
takes
into the stalk cell
migrates
In the young
place (Fig. 5).
and the second meiotic division
ascus,
ascus
nuclei
the first meiotic division
membranes around the nuclei (Figs,
Changes
in
lation
Fig.
—
After
content
ploidy
level
DNA
In
next
1500 Mb (then 1C
the DNA
content
DNA
division
=
1500
DNA
content
synthesis
was
and cell division
The C-value
can
:
(Figs. If, 4c).
2
=
In such nuclei, the
at
con-
the time
of sporu-
750 Mb, the
may
occur
same
1350 Mb for A. vivida
are
Figs, lg, 6b)
formed. In such
(Fig. 6c).
a
nuclei,
DNA
After Me2,
eight
DNA
content
was
paraphyses).
and mitosis (Mi;
mean
DNA
stable DNA content
lC-value like in
asynchronously (Fig. 3d).
mean
be measured exactly
mean
770 Mb and after Mi 810 Mb. Thereafter,
spores with the 2C-value
at
asc-
of the
infolding
of the nucleus has the 4C-value. The
(Fig. 6a).
After the second meiotic division (Me2;
ar
nuclei
eight
(Figs, lj, 4h).
after the first meiotic division (Me 1) when the formed nuclei have
following
form
If, 4c)
spore, the nucleus with
maturing
a
vacuoles
replication
determinedas 3060 Mb
(2C) until the
to
daughter
6
karyogamy
was
during
two
mitosis (Mi)
con-
karyogamy (K)
(Mel) (Figs.
four
to
ascospores results from the
li, 4f).
chromatin remains between
densing
by
rise
two
content
This binucleate cell,
mother cell, in which
(Me2) (Figs, lg, 4d) give
Formation of
(Figs, lh, 4e).
whereas the
The nucleus of the
respectively.
(Figs. 4a, 5).
with the basic DNA content, each of which divides
ospore
binucleate,
nuclei with the basic DNA
Fig. lh)
mean
the
nucle-
uninucleate asco-
content was
measured
108
Fig.
PERSOONIA
4.
Photomicrographs of fruit-body
normal
micrographs),
of the first meiotic
j.
nuclei
division;
postmeiotic mitosis;
with
a.
f.
d. nuclei
condensing chromatin, nucleus
nuclei
in
a
hypha;
k. nuclei
of,N. vivida
in croziers
young nuclei
in
a
of
Vol.
in
and
the
hair;
course
g.
asci;
a
1,
2002
f: fluorescence
b.
meiotic
endoreduplicationin
i.
micrographs;
zygote nucleus;
of the second
vacuoli;
1. nuclei in
Part
(a, b, d,
young
ascospores;
between
18,
Bar
=
10
nuclei
division;
spore;
giant mitochondria
paraphysis.
c.
h.
in the
µm.
c, e,
in the
e.
nuclei after
fertilizing
apex
g-1:
course
of
an
spore
ascus;
Kullman:
Nucleair
The spore nuclei ofNeottiella
species
in absence of mitotic cell
replication
not
DNA
arrested
at
the 2C-value
as
in
two Neottiella
studiedhere
division).
undergo endoreduplication (DNA
In N.
in N. vivida, but may
109
species
is
rutilans, endoreduplication
proceed
different rate in spores.
at a
In N. rutilans spores with DNA values of 2C, 3C, SC, and 6C have been measured and
endopolyploidy
can
5% and in N.
(tm ±
Ascogenous
hyphae
most
N.
is
form
to
a new
and the stalk cells, while the
All
uously (Fig. 5).
cell,
penultimate
karyogamy.
be
(Figs. 3a, b,
In N.
et
content
one
hyphae,
high
in
fungi
is known
Westergaard,
synthesis
is
DNA
et
are
1966)
are
a
not
to occur
contains
Aiton
content
with that
zygotes.
and
before and sometimes
usually
occurs
Neurospora
before karyogamy,
Shear & 8.0.
crassa
in all four nuclei of the crozier cells
shown) (Figs. 4a, 5).
general
visible in the
ascus
condition in
fungi
apex. The DNA
eight ascomycetes
multiple
al. (1996) examined the
Pelargonium zonale
compared
for nuclei in the initial
for formation of
It is
with
possible
that
homokaryotic
a
of
content
mito-
giant
(Fig. 4i).
was
found
N.F. de Vries)
to
to
be from 18.9
108 b.k.
( Bret-
Florenz.) (Weber, 1993). Obviously, the mitochondrionof N. vivida
content
found that DNA
of the Neottiella
1990).
mitochondriareveal the existence of
Kuroiwa
except
karyogamy
0Torulopsis glabrata (H.W.Anderson) Lodder &
with
case
after another from the terminal
determinedto be 60 Mb, 54 Mb, and 30 Mb
tanomyces custersii
both cells, the
case
nucleus from the terminal
cell initiates formationof new croziers contin-
used in
The mitochondrial genome size of
b.k.
nuclei
mitochondria
Giant mitochondria
was
a
asco-
synthesis
& Collins,
of giant
of
extremely large
4a, 5). However, in the
be formed
ascogenous
terminal and stalk cells
fruit-body (Bayman
migrate
vivida, premeiotic DNA replication
DNA
precise ontogeny
In this
repeatedly.
al., 1977), and synchronously
delayed premeiotic
chondria
branch
(Figs.
can
penultimate
replication
in N. rutilans (Rossen &
Dodge (Iyengar
asci
potentially
DNA
ofpremeiotic
Premeiotic DNA
DNA
may
crozier
nuclei of
can
be found within
can even
The
Neottiella vivida with
cell and the stalk cell (containing also the
type pattern of karyogamy,
as
study.
1981).
for this kind of research.
vivida, ascogenous hyphae
cell), elongate
after
x
13.0 pm ± 3%.
variation
Kimbrough,
difficultto
usually
appropriate species
penultimate
Time
x
two
23.1 |tm ± 5%
may reveal considerable variation in the pattern of
species
hyphal differentiation; significant
genus (!Thelebolus Tode: Fr.,
genous
In
22.8 pm ± 4%
at
at
hyphal differentiation
ascogenous
is the
sizes between the
in N. rutilans have been measured
vivida
There is evidence that
single
differentlevels. These uninucleate spores
to occur at
differences have been found in spore
studied: spore sizes
specimens
12.8
be assumed
No
heteroploid.
are
at
a
strands of DNA. Different DNA
development
of
during megasporogenesis
within the stacked
giant
and
mitochondria of the
megagametogenesis.
mitochondrion increased up
the megaspore mother cell stage;
contained 340-1700 Mb DNA.
contents
of
different number of strands in their nucleoids.
a
single
to
plant
They
40 times
stack of mitochondria
110
Fig.
PERSOONIA
5.
Ascogenous hyphal
nucleus
migrating
ascogenous
formation
from
hyphae, except
of
zygotes
in
differentiation.
the
terminal
for nuclei
Vol.
The
cell)
of the
asci. A. ascus; C.
Part
1,
penultimate
or
may
18,
2002
the stalk cell
elongate forming
new
initial, penultimate cell,
(containing
croziers.
can
be
All
also
the
nuclei
of
for
used
potentially
crozier.
Nuclear genome size
In this
of N.
study
the
genome
size of N. rutilans
(author's unpublished data)
sp.
reported
was
Genome sizes for Pezizales
vivida 750 Mb.
earlier for other
fungi
to
750 Mb for N.
determinedto be 530 Mb and that
from 12 Mb for Pulvinula
ranged
vivida. The
fall in this range
too
majority
of genome sizes
(Duran & Gray, 1989; Wittman-
Meixner, 1989;Zolan, 1995).
The overall genome size estimated in this
than that of the ascomycetous
cantly larger
data are available on internet:
the
Nagata, 1992) and 24
Mb
by
content
(Larraya
et
Mb (Kullman,
to
4.9 Mb
al.,
30 Mb (two
1989),
for
and other filamentous fungi. For
(Eidam)
crassa
subpopulations
in
31 Mb
21 Mb
one
signifi-
example,
(Sagawa
&
spore-print differing
in
ostreatus
(Peberdy
for Penicilliumpaxilli Bainier 23 Mb (Itoh
Podospora
Neurospora
was
cerevisiae (13 Mb,
et
al„
1993)
and 35
1999), for Trichophaea hemisphaerioides (Mouton) Graddon 23
2000),
nidulans
species
yeast Saccharomyces
(20%), Kullman, 2000),
plasma capsulatum Darling
gillus)
for Neottiella
genome sizes have been determined:for P.
following
DNA
ftp.ebi.ac.uk)
study
23
to
32 Mb
(Carr
& Shearer,
1998),
et
al., 1994), for Histo-
for Emericella (Asper-
Vuill. 26 Mb (Timberlake, 1978) to 31 Mb
anserina
Shear &
(Rabenh.)
Dodge
Niessl 34 Mb
39 Mb (Orbach,
(Brody
(Javerzat
1992) and 43
to
et
al.,
& Carbon,
1993),
for
45 Mb (Radford
Nucleair
DNA
in
two Neottiella
species
1
Kullman:
Fig.
6.
Histograms
Postfusion
meiotic
division
hypha in
G0/G2
zygote
cell
=
of the relative C-values
nuclei
(nuclei
in
up
2C);
cycle phase G0/G1
1C and G2M
=
2C;
(additionallight including
& Parish,
young
to
asci
c.
=
of nuclei in
(4C);
f. nuclei in
a
x
E.
typhina (Pers.)
=
2C;
of
cells,
stages
in
synthesis
in
e.
fruit-body (all
spores
nuclei
in
1999).
The last
two
life
a
on
largest
in
d. nuclei in
cell
a.
second
a
cycle phase
histograms)
and
noise
maximum).
hybrids Neotyphoidium
values represent the
of N. vivida.
after the
2C);
to
paraphysis
Schardl & M.R.
Tul. & C. Tul. and N.
cycle
nuclei
(nuclei up
Siegel
29 Mb, for
lolii Latch, M.Chr.
&
coenophialum (Morgan-Jones
W. Gams) Glenn, C.W. Bacon & Hanlin 55 Mb and 57 Mb,
al.,
in
ascus
measurements
i.e. the first
festucae Leuchtm.,
Epichloë (Fr.) Tul. & C. Tul. anamorphic
& Samuels
different
endoreduplication
1C and G2M
autofluorescence
1997), for Epichloë
b. DNA
respectively (Kuldau
genome sizes
reported
so
et
far for
filamentous fungi.
Ploidy
levels and
speciation
When among ascomycetes
(Weber, 1992), then, using
the
ploidy
level of N.
genome sizes, the
ploidy
rutilans is
level of N.
approximately
vivida
was
50
calculated
112
to
PERSOONIA
be 70. The
N. vivida.
species
two
which possess
form
Heteroploid
reticulate
a
Griffiths,
1999),
be
can
reticulum in N.
restricted
more
to
2002
distinguished only by
rutilans and delicate
warts
According
ornament.
cells of the
1,
Part
18,
of N. rutilans have
spores
tation. Some spores have
to
very similar and
are
incomplete
an
Vol.
a
of
high variability
which
Galitski
et
warts
connected
al. (1999) (see also Hieter &
ascomycetous yeast Saccharomyces cerevisiae,
differing
only
in their
ploidy,
gene
dosage,
but show different patterns of gene expression. In N. vivida and N. rutilans,
are
identical in
in
ornamen-
spore
completely
not
are
their spores,
regular
of DNA sequence information and relative
terms
the differences in spore ornamentation may be the result of different gene expressions
regulated by
ploidy-dependent
a
As the spores of N. rutilans
have the
spore
nuclei
tiales,
DNA
same
at
content.
gene.
are
heteroploid,
There is
yet
as
some
It is also known that
germination.
same
that in such
speculated
scribed
speciation,
related
will
polyploidy
suggested
as
and in spore width
respectively)
cases
&
by Bresinsky
content
Z.S.
(Fr.)
of their uniIt
(Weber, 1992).
confirm
repeatedly
of Leo-
species
and ‘H.’ rhodoleucus
genome size, differ only in the relative DNA
nucleate spores (1C and 2C,
of these
haploidization
closely
two
Dennis
‘Hymenoscyphus’ equisetinus (Velen.)
Bi, having the
spores of N. rutilans and N. vivida
evidence of
no
can
Wittmann-Bresinsky
be
de-
taxonomically
(1995) for
Boletales.
CONCLUSIONS
Within the
investigations
true
of Neottiella serve
fungi, species
due to theirextraordinarily
fluorescence. Neottiella vivida and N. rutilans
imately
750 Mb and 530 Mb,
different levels of
Probably
large variability
generation
cell is
verse
the
a
for metabolic
with
replicated
to
replication
of asci
asci),
one
hyphae,
be used in
two
karyogamy
for simultaneous
or
et
meiospore
formation. It
plastids ultimately produced
1967,1973).
as
The
appears
con-
to
be
al., 1974). This problem needs
hyphae
production
more
requires
principle maximally
production
regulation
of
of
frequent
a
in
species
further
large
with
study.
spore
i.e.
with small
large,
more
penultimate cells,
spore
case,
all
production.
production:
new
number of asci. It
species
economical. All
of zygotes. In this
be used for
eventually
can
formation of new croziers,
the second alternative occurs in
this argument
for the
intensive aerobic respiration
regulated by plastids,
for
alternatives allowing
after another,
the first alternative is
ever,
account
specimen.
endopolyploidy (Butterfass,
is
of spores has been
except for the nuclei in the initial, i.e.
DNA in ascogenous
However, there are
auto-
fungi.
nuclei of ascogenous
potential
cytogenetic
have genomes of approx-
associated with
The Neotiella-type pattern of karyogamy is in
have the
for
and low cell
in its spores
indicating
ascus,
activity
Chlamydomonas Ehrenberg (Blamire
further research in
to
objects
content
heteroploidy
that the number of
flowering plants
function of the level of
idea is that nuclear DNA
case
found
in which
of spore ornamentation within one
is demonstrated in
per
were
endopolyploidy
Giant mitochondria were found in the
and energy
excellent
respectively.
Neottiella rutilans is the first ascomycete
discovered.
as
nuclear DNA
large
formation
branches (instead of
can
be
primitive
speculated
that
ascomata, while
differentiatedascomata. How-
Kullman:
Nucleair
DNA
in
two Neottiella
113
species
ACKNOWLEDGEMENTS
The research
Nos.
3580
sultations
the
was
and
all-round
help.
Sincere thanks
manuscript.
I thank
are
Foundation. I thank Prof. A.
Dr. M.
and
(Deutsche Forschungsgemeinschaft)
the DFG
supported partly by
and 4989 of the Estonian Science
Rahi
due to Mrs. E.
and Dr.
for
Jaigma
A.
Raitviir
revising
Bresinsky
comments
for critical
the
by grants
for useful con-
text of the
English
on
manu-
script.
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